Rhamnose synthase derived from stevia and gene

Patent metadata

Field	Value
Publication number	US-11447765-B2
Application number	US-201816604303-A
Country	US
Kind code	B2
Filing date	Apr 11, 2018
Priority date	Apr 12, 2017
Publication date	Sep 20, 2022
Grant date	Sep 20, 2022

Abstract

The purpose of the present invention is to provide a protein, said protein having an activity of synthesizing rhamnose from glucose, and a polynucleotide encoding the same. Provided are a rhamnose synthase derived from stevia and a method for producing rhamnose from glucose using a gene. Also provided is a method for producing a steviol glycoside using the rhamnose synthase derived from stevia.

First claim

The invention claimed is:

1. A method comprising reacting, outside of a stevia plant, glucose and the protein according to any one selected from the group consisting of the following (a) to (c) to produce rhamnose:
(a) a protein consisting of the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, or 10;
(b) a protein consisting of an amino acid sequence wherein 1 to 13 amino acids are deleted, substituted, inserted, or added in the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, or 10 and having an activity to produce rhamnose from glucose;
(c) a protein having an amino acid sequence having a sequence identity of 98% or more to the amino acid sequence of SEQ ID NO: 2, 4, 6, 8, or 10 and having an activity to produce rhamnose from glucose.

2. The method according to claim 1 , wherein:
the glucose is in the form of uridine diphosphate glucose, or
the rhamnose is in the form of uridine diphosphate rhamnose.

3. The method of claim 1 , further comprising:
transferring the rhamnose to steviol or a steviol glycoside to produce steviol glycoside.

4. The method according to claim 3 , wherein the rhamnose is in the form of uridine diphosphate rhamnose.

5. The method according to claim 3 , wherein the transferring of the rhamnose to steviol or a steviol glycoside is performed by using: a host cell comprising (i) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 11 or 13; or (ii) a polynucleotide encoding a protein having an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence of SEQ ID NO: 12 or 14 and having an activity to add rhamnose to glucose at position 13 in the steviol glycoside via a 1→2 bond.

6. The method according to claim 5 , wherein the host cell comprises at least one polynucleotide of the following (a) to (e):
(a) (i) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 15; or (ii) a polynucleotide encoding a protein having an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence of SEQ ID NO: 16 and having an activity to add glucose to the hydroxyl group at position 13 in a steviol glycoside;
(b) (i) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 17; or (ii) a polynucleotide encoding a protein having an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence of SEQ ID NO: 18 and having an activity to add glucose to the carboxylic acid at position 19 in a steviol glycoside;
(c) (i) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 19; or (ii) a polynucleotide encoding a protein having an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence of SEQ ID NO: 20 and having an activity to add glucose at position 3 of the glucose at position 13 in a steviol glycoside via a 1→3 bond;
(d) (i) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 11 or 13; or (ii) a polynucleotide encoding a protein having an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence of SEQ ID NO: 12 or 14 and having an activity to add glucose to the glucose at position 19 in a steviol glycoside via a 1→2 bond;
(e) (i) a polynucleotide comprising the nucleotide sequence of SEQ ID NO: 19; or (ii) a polynucleotide encoding a protein having an amino acid sequence having a sequence identity of 90% or more to the amino acid sequence of SEQ ID NO: 20 and having an activity to add glucose to the glucose at position 19 in a steviol glycoside via a 1→3 bond.

7. The method of claim 1 , further comprising:
adding the produced rhamnose to a raw material of the sweetener, the food or beverage, or the pharmaceutical product; and
preparing the sweetener, the food or beverage, or the pharmaceutical product.

8. The method according to claim 7 , wherein the rhamnose is in the form of uridine diphosphate rhamnose.

Assignees

• Suntory Holdings Ltd

Inventors

• Ono Eiichiro
• Ochiai Misa
• Iwaki Kazunari
• HIRAI Tadayoshi

Classifications

• A23L27/30

  Artificial sweetening agents · CPC title

• A23L33/13

  Nucleic acids or derivatives thereof (A23L33/145 takes precedence) · CPC title

• A23L2/66

  Proteins · CPC title

• C12P19/56

  having an oxygen atom of the saccharide radical directly bound to a condensed ring system having three or more carbocyclic rings, e.g. daunomycin, adriamycin · CPC title

• C12Q2600/13

  Plant traits · CPC title